CN111226694A - Method for cultivating and obtaining cordyceps militaris sporocarp by taking tenebrio molitor larvae as raw materials - Google Patents
Method for cultivating and obtaining cordyceps militaris sporocarp by taking tenebrio molitor larvae as raw materials Download PDFInfo
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Abstract
A method for culturing Cordyceps militaris fruiting body with Tenebrio molitor larva as raw material comprises inoculating Cordyceps militaris mother strain in liquid culture medium, and culturing in shaking table at full temperature to obtain liquid strain; the formula of the liquid culture medium comprises 18-25% of potato, 1-4% of glucose and KH2PO40.05%~0.2%、MgSO4·7H20.05-0.2% of O, 0.2-0.5% of peptone and the balance of water; adding tenebrio molitor larvae into the cultivation bottle, adding a glucose solution with the pH =8 and the concentration of 1%, sealing, and sterilizing at high temperature and high pressure to obtain a tenebrio molitor cultivation medium; uniformly spraying liquid strains on the surface of a tenebrio molitor culture medium; placing the culture bottle on a shelf of a culture room for culturing, collecting before the middle-upper part of the cordyceps militaris sporocarp is densely covered with ascocarps but not scattered with spores, and separating the cordyceps cake from the cordyceps militaris sporocarp to obtain the cordyceps militaris sporocarp. Simplified process, easy industrialization, and yellow mealworm larva as main cultivation materialThe obtained cordyceps militaris sporocarp is rich in ascocarp and ascospore, and the contents of cordycepin, cordyceps polysaccharide, SOD enzyme and other nutrient components are higher.
Description
Technical Field
The invention relates to the field of edible fungus cultivation, in particular to a method for obtaining cordyceps militaris sporocarp by cultivating yellow mealworm larvae as a raw material.
Background
Cordyceps militaris is also called as Cordyceps militaris, belonging to the genus Cordyceps of Clavipitaceae of Hypocreales of Ascomycetes, and is a model species of Cordyceps, wherein the stroma and pupa are formed by parasitizing Cordyceps militaris fungi on pupa of Lepidoptera insects. Because the main active ingredients of the cordyceps sinensis are similar to or higher than that of the cordyceps sinensis and mature culture conditions exist, the cordyceps sinensis is an effective substitute of the cordyceps sinensis.
In the prior art, the cordyceps militaris fruiting body using yellow mealworm pupa as a host and the cultivation method thereof disclosed by CN100392063C have the disadvantages that the yellow mealworm pupa is a host, the injection and inoculation are inconvenient to operate and are not beneficial to industrial production, and the fruiting body is less; CN103688760A discloses a method for culturing artificial cordyceps sinensis by taking yellow mealworms as carriers, which is inconvenient to operate by injection inoculation, mainly takes the harvesting of the zoophora, and has small inoculation amount, so that no fruiting body is generated or the cordyceps sinensis is extremely short; CN103749149A discloses a method for culturing artificial cordyceps sinensis by taking yellow mealworm pupae as a carrier, which takes the pupae as a host, only uses alcohol for surface disinfection and then inoculates in an injection mode, the pupae is in risk of carrying pathogenic bacteria, and the sporocarp is short and small; CN105493899B discloses a method for culturing a cordyceps product by taking yellow mealworm pupae as a carrier, which takes the yellow mealworm pupae as a host and performs injection inoculation to obtain the product which is a cordyceps without sporophytes; CN108184540A discloses a method for cultivating cordyceps militaris by using live tenebrio molitor larvae as hosts, wherein the obtained fruiting bodies are 1.8-3cm, and an injection inoculation mode is adopted.
In the related prior art, the mycelium of the cordyceps militaris is obtained simply by adopting injection and infection, on one hand, the operation is complicated, the biological conversion efficiency is not ideal, the obtained entity nutrient content is deficient or not good, for example, no sporophore or few sporophore exists, and the sporophore is only 1.8-3cm, on the other hand, the existing treatment mode is not beneficial to industrial production.
Disclosure of Invention
The method for cultivating and obtaining the cordyceps militaris sporocarp by taking the tenebrio molitor larvae as the raw materials is mainly used for overcoming the defects of the related prior art, the process is simplified, the operability is improved, the large-scale and industrial production is favorably realized, the tenebrio molitor larvae are used for replacing rice and wheat as the main cultivation raw materials, the ascospores and ascospores of the cordyceps militaris sporocarp obtained by cultivation are rich, the contents of nutrient components such as cordycepin, cordyceps polysaccharide, SOD enzyme and the like are higher, the medicinal components of the cordyceps militaris sporocarp are improved, and the cordyceps militaris sporocarp has high food and medicinal values.
In order to achieve the above object, the present invention employs the following techniques:
a method for cultivating and obtaining cordyceps militaris sporocarp by taking yellow mealworm larvae as a raw material is characterized by comprising the following steps:
inoculating the cordyceps militaris mother strain to a liquid culture medium, and performing shaking table culture at full temperature to obtain a liquid strain with uniform bacterium ball size, viscous bacterium liquid, wall-hanging property and no turbidity; wherein the liquid culture medium is prepared from the following raw materials in parts by weight: peeled potato 18% -25%, glucose 1% -4% and KH2PO40.05%~0.2%、MgSO4·7H20.05-0.2% of O, 0.2-0.5% of peptone and the balance of water;
adding tenebrio molitor larvae into a cultivation bottle, adding a glucose solution with the pH =8 and the concentration of 1%, sealing with an OPP sealing film and a leather sheath, and sterilizing at 120-125 ℃ and 0.10-0.15 MPa for 1-2 h to obtain a tenebrio molitor cultivation medium;
uniformly spraying the liquid strain diluted by sterile water on the surface of a tenebrio molitor culture medium;
placing the cultivation bottle on a shelf of a culture room for hypha cultivation, wherein the cultivation temperature is 18-22 ℃, and hypha covers are 8-10 d;
the relative humidity of air is increased to 80-90%, and the cultivation bottle is subjected to light supplement treatment, wherein the illumination intensity is 800-1100 Lux, and the light supplement is performed for 12 hours each day;
when cordyceps militaris fruiting body primordium appears on the surface of the tenebrio molitor culture medium, cutting a small opening on the sealing film, and entering a fruiting body growth stage;
in the growth stage of the sporocarps, the temperature is 18-22 ℃, the relative humidity of air is 80-90%, and the cultivation room is ventilated for 2 times every day, wherein each time lasts for 0.5-2 hours;
collecting the fruiting bodies of Cordyceps militaris before the fruiting bodies of Cordyceps militaris are densely covered with ascocarpes but not scattered with spores, separating the Cordyceps militaris cakes from the fruiting bodies of Cordyceps militaris during collection, respectively drying at 40-60 deg.C, sealing, keeping out of the sun, and storing in a moisture-proof manner to obtain the fruiting bodies of Cordyceps militaris.
Further, the cordyceps militaris mother strain is obtained by collecting mature wild cordyceps militaris individuals without scattering spores, performing tissue separation in a laboratory, and screening.
Further, the preparation of the liquid culture medium comprises the steps of weighing peeled potatoes, cutting the peeled potatoes into slices of 2-3 mm, adding the slices into boiling water at the temperature of 100-120 ℃, boiling for 15-20 minutes until the peeled potatoes are crisp but not soft; filtering with 6-8 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are filled into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized at the temperature of 120-125 ℃ and the pressure of 0.10-0.15 MPa for 0.5-1 h to prepare the medical composition. In the preparation process, the water reduced by evaporation needs to be supplemented, and the water supplementing amount is determined according to the constant volume matched with the total amount of the liquid culture medium. Take a total amount of 1000mL of liquid medium formula as an example: peeled potato 200 g (20%), glucose 20g (2%), KH2PO41 g (0.1%), MgSO4·7H2And (3) 1 g (0.1%) of O, 3 g (0.3%) of peptone and the balance of water, wherein the evaporation of water is reduced in the boiling process, and finally, water is supplemented to a constant volume of 1000mL, subpackaging is carried out, and the sterilization is carried out at 120-125 ℃ and 0.10-0.13 MPa for 0.5-1 h.
Further, parameters of the full-temperature shaking table are 18-25 ℃ and 120-160 r/min, and the cultivation lasts for 3-5 d.
Further, when the culture bottle has a volume of 700-900 mL, 18-25 g of the tenebrio molitor larvae are added, and 10-20 mL of a 1% glucose solution with pH =8 is added.
Further, when the liquid strain diluted by sterile water is uniformly sprayed on the surface of the tenebrio molitor cultivation medium, 5-6 mL of the liquid strain is used per bottle.
The invention has the beneficial effects that:
1. the method is mainly characterized in that yellow mealworm larvae are used as main cultivation raw materials, mycelium infecting cordyceps militaris is not simply obtained, and the purpose of obtaining cordyceps militaris sporocarp through cultivation and growth is mainly achieved;
2. the tenebrio molitor larvae are used for replacing rice and wheat as main cultivation raw materials, ascospores and ascospores of fruiting bodies of cordyceps militaris obtained by cultivation are rich, and the contents of nutrient components such as cordycepin, cordyceps polysaccharide, SOD (superoxide dismutase) and the like are higher, so that the medicinal components of the fruiting bodies of cordyceps militaris are improved, and the cordyceps militaris has high food and medicinal values;
3. about 9 g-16 g of cordyceps militaris fruiting bodies can be obtained by cultivating 18 g-25 g of tenebrio molitor larvae, the cultivation growth period is 70 d-75 d, and the biological conversion rate is basically 50% -72%.
4. In the prior art, the purpose of harvesting the zoophobous is to obtain small and short sporophores without sporophores or with few sporophores; meanwhile, the prior art mostly adopts injection type inoculation, which is inconvenient to operate; the treatment mode of the embodiment of the scheme is that tenebrio molitor larvae are used as main materials of the culture medium, sporocarp with the length of 10-15 cm can be obtained by improving the strain formula and the inoculation mode, the sporocarp is 5-10 times that of the prior art, and the scheme is more favorable for industrial production and development and is a more optimal choice for promoting large-scale production.
Drawings
FIG. 1 is a diagram illustrating the cultivation of the Tenebrio molitor from the cultivation medium to the primordia of the fruiting body of Cordyceps militaris in the first embodiment.
FIG. 2 is a diagram showing a plant fruit body cultivated according to the first embodiment.
FIG. 3 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
FIG. 4 is a diagram showing the actual plants of the grass carpophores cultivated in example two.
FIG. 5 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
FIG. 6 is a diagram showing the actual plants of the grass carpophores cultivated in the third example.
FIG. 7 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
Detailed Description
Example one
1. Obtaining a cordyceps militaris mother strain: collecting mature wild cordyceps militaris individuals without scattering spores from the afterveins of the Changbai mountain in northeast China, carrying out tissue separation in a laboratory, and screening to obtain high-quality cordyceps militaris mother strain.
2. Preparing a liquid culture medium: the culture medium comprises peeled potato 18%, glucose 2%, and KH2PO40.05%、MgSO4·7H20.05% of O, 0.4% of peptone and the balance of water. During the preparation process, the water reduced by evaporation needs to be supplemented, and the water supplement amount is matched with the total amount of the liquid culture medium.
Weighing peeled potato, cutting into 2mm slices, decocting in 120 deg.C boiling water for 15 min, filtering with 6 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are subpackaged into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized at 122 ℃ and 0.12MPa for 0.5h to prepare the oral liquid for later use.
3. Liquid strain propagation: screening high-quality Cordyceps militaris mother strain, inoculating to liquid culture medium, and culturing in a full-temperature shaking table with parameters of 18 deg.C and 160r/min for 5 days to obtain high-quality liquid strain with uniform pellet size, viscous strain liquid, and no turbidity.
4. Preparing a culture medium for cultivating yellow mealworms: adding 18g of tenebrio molitor larvae and 10mL of 1% glucose solution with pH =8 into a cultivation bottle with a volume of 700mL, sealing with a high pressure resistant OPP sealing film and a leather sheath, and sterilizing at 122 ℃ and 0.13MPa for 1h to obtain the tenebrio molitor cultivation medium.
5. Inoculation: and uniformly spraying the liquid strains diluted by sterile water on the surface of the tenebrio molitor culture medium on an indoor superclean workbench, wherein each bottle uses about 5mL of the liquid strains.
6. Cultivation:
inoculating a tenebrio molitor larva culture medium, and placing a culture bottle on a shelf of a culture room for mycelium culture, wherein the culture temperature is 18 ℃, and the mycelium cover needs about 8 days;
increasing the relative humidity of air to 80%, and performing light supplement treatment on the cultivation bottle by using a fluorescent lamp, wherein the illumination intensity is 800Lux, and the light supplement is performed for 12 hours every day to promote the color conversion of the tenebrio molitor larva mycelium and the differentiation of fruiting body primordium;
around 25 days, cordyceps militaris sporophore primordium appears on the surface of the tenebrio molitor culture medium, a small opening is cut on the sealing film by a wallpaper blade, and the step of sporophore growth management is carried out; a real object/scene diagram of the implementation process is shown in fig. 1;
the temperature of the fruiting body in the growth stage is 18 ℃, the relative humidity of air is 80%, and the cultivation room is ventilated for 2 times every day, 2 hours each time.
7. Harvesting: collecting fruiting body of Cordyceps militaris after fruiting body grows for about 37 days, densely distributing ascocarp at middle upper part of the fruiting body of Cordyceps militaris, but before spore scattering, separating Cordyceps cake and fruiting body of Cordyceps militaris, oven drying at 40 deg.C respectively, sealing, keeping out of the sun, and keeping out of the sun.
FIG. 2 is a diagram of a plant fruit body cultivated according to the first embodiment.
FIG. 3 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
In this example, the cultivation growth period totals about 70d, corresponding to the partial test results as given in the following table:
example two
1. Obtaining a cordyceps militaris mother strain: collecting mature wild cordyceps militaris individuals without scattering spores from the afterveins of the Changbai mountain in northeast China, carrying out tissue separation in a laboratory, and screening to obtain high-quality cordyceps militaris mother strain.
2. Preparing a liquid culture medium: the culture medium comprises peeled potato 25%, glucose 4%, and KH2PO40.2%、MgSO4·7H20.2% of O, 0.5% of peptone and the balance of water. During the preparation process, the water reduced by evaporation needs to be supplemented, and the water supplement amount is matched with the total amount of the liquid culture medium.
Weighing peeled potato, cutting into 3mm slices, decocting in 100 deg.C boiling water for 18 min, filtering with 8 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are separately filled into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized for 1h at the temperature of 120 ℃ and under the pressure of 0.15MPa to obtain the product for later use.
3. Liquid strain propagation: screening high-quality Cordyceps militaris mother strain, inoculating to liquid culture medium, and culturing in a full-temperature shaking table with parameters of 25 deg.C and 120r/min for 3d to obtain high-quality liquid strain with uniform pellet size, viscous strain liquid, and no turbidity.
4. Preparing a culture medium for cultivating yellow mealworms: adding 25g of tenebrio molitor larvae into a culture bottle with a volume of 900mL, adding 20mL of glucose solution with pH =8 and a concentration of 1%, sealing with a high pressure resistant OPP sealing film and a leather sheath, and sterilizing at 125 ℃ and 0.10MPa for 2h to obtain the tenebrio molitor culture medium.
5. Inoculation: and uniformly spraying the liquid strains diluted by sterile water on the surface of the tenebrio molitor culture medium on an indoor superclean workbench, wherein each bottle uses about 6mL of the liquid strains.
6. Cultivation:
inoculating a tenebrio molitor larva culture medium, and placing a culture bottle on a shelf of a culture room for hypha culture, wherein the culture temperature is 22 ℃, and the hypha cover needs 10 days;
increasing the relative humidity of air to 90%, and performing light supplement treatment on the cultivation bottle by using a fluorescent lamp, wherein the illumination intensity is 1100Lux, and the light supplement is performed for 12 hours every day to promote the color conversion of the tenebrio molitor larva mycelium and the differentiation of fruiting body primordium;
around 30 days, cordyceps militaris sporophore primordium appears on the surface of the tenebrio molitor culture medium, a small opening is cut on the sealing film by a wallpaper blade, and the step of sporophore growth management is carried out;
the temperature of the fruiting body in the growth stage is 22 deg.C, the relative humidity of air is 90%, and the cultivation room is ventilated for 0.5 hr for 2 times per day.
7. Harvesting: collecting fruiting body of Cordyceps militaris after fruiting body grows for about 35 days, densely distributing ascocarp at middle upper part of the fruiting body of Cordyceps militaris, but before spore scattering, separating Cordyceps cake and fruiting body of Cordyceps militaris, oven drying at 60 deg.C respectively, sealing, keeping out of the sun, and keeping out of the sun.
FIG. 4 is a diagram showing a plant fruit body cultivated in the second embodiment.
FIG. 5 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
In this example, the cultivation growth period totals about 75d, corresponding to the partial test results as given in the following table:
EXAMPLE III
1. Obtaining a cordyceps militaris mother strain: collecting mature wild cordyceps militaris individuals without scattering spores from the afterveins of the Changbai mountain in northeast China, carrying out tissue separation in a laboratory, and screening to obtain high-quality cordyceps militaris mother strain.
2. Preparing a liquid culture medium: the culture medium comprises peeled potato 20%, glucose 1%, and KH2PO40.15%、MgSO4·7H20.15% of O, 0.3% of peptone and the balance of water. During the preparation process, the water reduced by evaporation needs to be supplemented, and the water supplement amount is matched with the total amount of the liquid culture medium.
Weighing peeled potato, cutting into 3mm slices, adding into 110 deg.C boiling water, decocting for 20 min, filtering with 8 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are filled into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized at 125 ℃ and 0.10MPa for 0.75h to prepare the product for later use.
3. Liquid strain propagation: screening high-quality Cordyceps militaris mother strain, inoculating to liquid culture medium, and culturing in a full-temperature shaking table with parameters of 21 deg.C and 130r/min for 4 days to obtain high-quality liquid strain with uniform pellet size, viscous strain liquid, and no turbidity.
4. Preparing a culture medium for cultivating yellow mealworms: adding 20g of tenebrio molitor larvae and 15mL of 1% glucose solution with pH =8 into a cultivation bottle with a capacity of 750mL, sealing with a high pressure resistant OPP sealing film and a leather sheath, and sterilizing at 124 ℃ and 0.15MPa for 1.2h to obtain the tenebrio molitor cultivation medium.
5. Inoculation: and uniformly spraying the liquid strains diluted by sterile water on the surface of the tenebrio molitor culture medium on an indoor superclean workbench, wherein each bottle uses about 6mL of the liquid strains.
6. Cultivation:
inoculating a tenebrio molitor larva culture medium, and placing a culture bottle on a shelf of a culture room for hypha culture, wherein the culture temperature is 20 ℃, and the hypha cover needs 10 days;
increasing the relative humidity of air to 85%, and performing light supplement treatment on the cultivation bottle by using a fluorescent lamp, wherein the illumination intensity is 1000Lux, and the light supplement is performed for 12 hours every day to promote the color conversion of the tenebrio molitor larva mycelium and the differentiation of the fruiting body primordium;
marking a small opening on the sealing film by a wallpaper blade when cordyceps militaris sporocarp primordium appears on the surface of the tenebrio molitor culture medium for about 27 days, and entering a sporocarp growth management stage;
the temperature of the fruiting body in the growth stage is 21 deg.C, the relative humidity of air is 82%, and the cultivation room is ventilated for 1 hr 2 times daily.
7. Harvesting: collecting fruiting body of Cordyceps militaris after fruiting body grows for about 36 days, densely distributing ascocarp at middle upper part of the fruiting body of Cordyceps militaris, but before spore scattering, separating Cordyceps cake and fruiting body of Cordyceps militaris, oven drying at 50 deg.C respectively, sealing, keeping out of the sun, and keeping out of the sun.
FIG. 6 is a schematic diagram of the fruiting body of the second embodiment.
FIG. 7 is a diagram of the separated insect cake and the fruiting body of Cordyceps militaris in the example.
In this example, the cultivation growth period totals about 73 days, corresponding to the partial test results as given in the following table:
example four
1. Obtaining a cordyceps militaris mother strain: collecting mature wild cordyceps militaris individuals without scattering spores from the afterveins of the Changbai mountain in northeast China, carrying out tissue separation in a laboratory, and screening to obtain high-quality cordyceps militaris mother strain.
2. Preparing a liquid culture medium: the culture medium comprises peeled potato 21%, glucose 3%, and KH2PO40.1%、MgSO4·7H20.1% of O, 0.2% of peptone and the balance of water. During the preparation process, the water reduced by evaporation needs to be supplemented, and the water supplement amount is matched with the total amount of the liquid culture medium.
Weighing peeled potato, cutting into 2mm slices, decocting in 100 deg.C boiling water for 15 min, filtering with 7 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are subpackaged into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized at 124 ℃ and 0.13MPa for 0.6h to prepare the oral liquid for later use.
3. Liquid strain propagation: screening high-quality cordyceps militaris mother strain, inoculating the strain to a liquid culture medium, and culturing for 4d by using a full-temperature shaking table with parameters of 20 ℃ and 140r/min to obtain high-quality liquid strain with uniform bacterium ball size, viscous bacterium liquid and wall-hanging property and no turbidity.
4. Preparing a culture medium for cultivating yellow mealworms: 22g of tenebrio molitor larvae and 18mL of 1% glucose solution with pH =8 were added to a 800 mL-volume culture flask, and the flask was sealed with a high pressure resistant OPP sealing film and a leather sheath, and sterilized at 120 ℃ and 0.13MPa for 1.5 hours to obtain a tenebrio molitor culture medium.
5. Inoculation: and uniformly spraying the liquid strain diluted by sterile water on the surface of the tenebrio molitor culture medium on an indoor super-clean workbench for inoculation, wherein each bottle uses about 5.5mL of the liquid strain.
6. Cultivation:
inoculating a tenebrio molitor larva culture medium, and placing a culture bottle on a shelf of a culture room for hypha culture at the culture temperature of 20 ℃ for about 9 days for hypha cover;
increasing the relative humidity of air to 85%, and performing light supplement treatment on the cultivation bottle by using a fluorescent lamp, wherein the illumination intensity is 1000Lux, and the light supplement is performed for 12 hours every day to promote the color conversion of the tenebrio molitor larva mycelium and the differentiation of the fruiting body primordium;
about 28 days, cordyceps militaris sporophore primordium appears on the surface of the tenebrio molitor culture medium, a small opening is cut on the sealing film by a wallpaper blade, and the step of sporophore growth management is carried out;
the temperature of the fruiting body in the growth stage is 20 deg.C, the relative humidity of air is 85%, and the cultivation room is ventilated for 1 hr for 2 times per day.
7. Harvesting: collecting fruiting body of Cordyceps militaris after fruiting body grows for about 34 days, densely distributing ascocarp at middle upper part of the fruiting body of Cordyceps militaris, but before spore scattering, separating Cordyceps cake and fruiting body of Cordyceps militaris, oven drying at 50 deg.C respectively, sealing, keeping out of the sun, and keeping out of the sun.
In this example, the cultivation growth period totals about 71d, corresponding to the partial test results as given in the following table:
according to the embodiment of the application, 18 g-25 g of tenebrio molitor larvae are used for obtaining about 9 g-16 g of cordyceps militaris sporocarp through cultivation, the cultivation growth period is 70 d-75 d, and the biological conversion rate is basically 50% -72%. The fruiting body and ascospores of Cordyceps militaris obtained by the method are rich, and the contents of nutrients such as cordycepin, Cordyceps polysaccharide and SOD enzyme are higher, so that the medicinal components of the fruiting body of Cordyceps militaris are improved, and the medicinal value is higher.
120g of cordyceps sinensis fruiting bodies obtained in the first embodiment are subjected to sample feeding detection, analysis and detection are carried out according to relevant standards such as GB 5009.93-2010, GB5413.13-2010, GB/T5009.171-2003, GB/T5009.84-2003, GB/T5009.92-2003 and GB/T5009.82-2003, and the detection results are shown in the following table:
180g of cordyceps sinensis fruiting bodies obtained in the mode of the third embodiment are subjected to sample feeding detection, and are analyzed and detected according to the standards of GB 5009.93-2010, GB5413.13-2010, GB/T5009.171-2003, GB/T5009.84-2003, GB/T5009.92-2003, GB/T5009.82-2003 and the like, and the detection results are shown in the following table:
the cultivation and production technology provided by the embodiment of the application comprises the cordyceps militaris strain preparation and management key points of each cultivation link, is a novel technical method for obtaining cordyceps militaris fruiting bodies, is popular and easy to understand, has remarkable effect, can be modeled in production process, and is favorable for industrial cultivation and production.
The above examples are merely for illustrating the technical means of the present application, but the technical means of the present application should not be limited to the above examples.
Claims (8)
1. A method for cultivating and obtaining cordyceps militaris sporocarp by taking yellow mealworm larvae as a raw material is characterized by comprising the following steps:
inoculating the cordyceps militaris mother strain to a liquid culture medium, and performing shaking table culture at full temperature to obtain a liquid strain with uniform bacterium ball size, viscous bacterium liquid, wall-hanging property and no turbidity; wherein the liquid culture medium is prepared from the following raw materials in parts by weight: peeled potato 18% -25%, glucose 1% -4% and KH2PO40.05%~0.2%、MgSO4·7H20.05-0.2% of O, 0.2-0.5% of peptone and the balance of water;
adding tenebrio molitor larvae into a cultivation bottle, adding a glucose solution with the pH =8 and the concentration of 1%, sealing with an OPP sealing film and a leather sheath, and sterilizing at 120-125 ℃ and 0.10-0.15 MPa for 1-2 h to obtain a tenebrio molitor cultivation medium;
uniformly spraying the liquid strain diluted by sterile water on the surface of a tenebrio molitor culture medium;
placing the cultivation bottle on a shelf of a culture room for hypha cultivation, wherein the cultivation temperature is 18-22 ℃, and hypha covers are 8-10 d;
the relative humidity of air is increased to 80% -90%, the cultivation bottle is subjected to light supplement treatment, the illumination intensity is 800-1100 Lux, and light supplement is carried out for 12 hours each day;
when cordyceps militaris fruiting body primordium appears on the surface of the tenebrio molitor culture medium, cutting a small opening on the sealing film, and entering a fruiting body growth stage;
the sporophore growth stage is carried out at the temperature of 18-22 ℃, the relative air humidity is 80-90%, and the cultivation room is ventilated for 2 times every day, wherein each time lasts for 0.5-2 hours;
collecting the fruiting bodies of Cordyceps militaris before the fruiting bodies of Cordyceps militaris are densely covered with ascocarpes but not scattered with spores, separating the Cordyceps militaris cakes from the fruiting bodies of Cordyceps militaris during collection, respectively drying at 40-60 deg.C, sealing, keeping out of the sun, and storing in a moisture-proof manner to obtain the fruiting bodies of Cordyceps militaris.
2. The method for cultivating and obtaining cordyceps militaris fruiting bodies using tenebrio molitor larvae as raw materials according to claim 1, wherein the cordyceps militaris mother strain is obtained by collecting mature wild cordyceps militaris individuals without scattering spores, performing tissue separation in a laboratory, and screening.
3. The method for cultivating and obtaining the cordyceps militaris sporocarp by taking the tenebrio molitor larvae as the raw material according to claim 1, wherein the liquid culture medium is prepared by weighing peeled potatoes, cutting the peeled potatoes into slices with the size of 2-3 mm, adding the slices into boiling water at the temperature of 100-120 ℃, boiling the slices for 15-20 minutes until the slices are crisp but not rotten; filtering with 6-8 layers of gauze, adding glucose and KH into the filtrate2PO4、MgSO4·7H2O, peptone and water are filled into a small infusion bottle with the capacity of 250mL, the filling amount is 80 mL/bottle, and the small infusion bottle is sterilized at the temperature of 120-125 ℃ and the pressure of 0.10-0.15 MPa for 0.5-1 h to prepare the medical composition.
4. The method for cultivating and obtaining cordyceps militaris sporocarp by taking yellow mealworm larvae as raw materials according to claim 3, wherein in the preparation of the liquid culture medium, water reduced by evaporation needs to be supplemented, and the water supplementing amount is determined according to constant volume until the water supplementing amount is matched with the total amount of the liquid culture medium.
5. The method for cultivating and obtaining cordyceps militaris sporocarp by taking yellow mealworm larvae as raw materials according to claim 1, wherein parameters of a full-temperature shaking table are 18-25 ℃ and 120-160 r/min, and the cordyceps militaris sporocarp is cultivated for 3-5 days.
6. The method for cultivating and obtaining cordyceps militaris fruiting body using yellow mealworm larvae as raw material according to claim 1, wherein when the cultivating bottle has a volume of 700-900 mL, 18-25 g of yellow mealworm larvae are added, and 10-20 mL of 1% glucose solution with pH =8 is added.
7. The method for cultivating and obtaining cordyceps militaris fruiting bodies using tenebrio molitor larvae as raw materials according to claim 6, wherein 5-6 mL of liquid spawn is used per bottle when the liquid spawn diluted with sterile water is uniformly sprayed on the surface of a culture medium for cultivating the tenebrio molitor.
8. A cordyceps militaris fruiting body, which is obtained by the method for obtaining the cordyceps militaris fruiting body by cultivating the yellow mealworm larvae as the raw material according to any one of claims 1 to 7.
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